title: Improvements in transfection efficiency with chitosan modified poly(DL-lactide-co-glycolide) nanospheres prepared by the emulsion solvent diffusion method, for gene delivery.
abstract: This study sought to evaluate the in vitro transfection efficiency of plasmid DNA (pDNA)-loaded chitosan-modified poly(DL-lactide-co-glycolide) nanospheres (CS-PLGA NS) in a gene-delivery system. Using the emulsion solvent diffusion (ESD) method, pDNA-loaded PLGA NS was prepared and the surface of the PLGA NS was modified by binding to CS. Gene transfection ability of CS-PLGA NS was examined in A549 cells. The luciferase gene was used as a reporter gene. The pattern of luciferase activity by pDNA-loaded CS-PLGA NS was initially weak, but gradually grew stronger before decreasing activity. These phenomena should be in accordance with the sustained-release profile of pDNA from PLGA NS in the cytosol and the pDNA protection against DNase. Positively charged CS-PLGA NS was found, by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay, not to exhibit cytotoxicity on A549 cells. These results suggest that CS-PLGA NS are potential contributors to efficient pDNA delivery due to their increased interactions with cells and lack of cytotoxic effects.
authors.affiliation: Laboratory of Pharmaceutical Engineering, School of Pharmacy, Aichi Gakuin University, Chikusa, Nagoya, Japan.
authors.forename: Kohei
authors.initials: K
authors.lastname: Tahara
authors.forename: Takeshi
authors.initials: T
authors.lastname: Sakai
authors.forename: Hiromitsu
authors.initials: H
authors.lastname: Yamamoto
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Takeuchi
authors.forename: Naohide
authors.initials: N
authors.lastname: Hirashima
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Kawashima
mesh_terms: D045744:Cell Line, Tumor; D048271:Chitosan / Q000737:chemistry*; D004058:Diffusion; D004655:Emulsions / Q000737:chemistry; D006801:Humans; D019344:Lactic Acid / Q000008:administration & dosage / Q000737:chemistry* / Q000633:toxicity; D054118:Nanospheres / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011100:Polyglycolic Acid / Q000008:administration & dosage / Q000737:chemistry* / Q000633:toxicity; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D012997:Solvents / Q000737:chemistry; D014162:Transfection / Q000379:methods
chemical_list: D004655:Emulsions; D012997:Solvents; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D048271:Chitosan